Tomato Culture

Purpose: To initiate cultures of tomato using various plant parts and culture media. Some of the tissue will be used for subsequent transformations.

 

1.5 liter equivalent
15 ml    MS FeEDTA  add first
15 ml    MS Sulfates
15 ml    Halides
15 ml    P, B, Mo
15 ml    B5 Vitamins
2.5 g     NH4NO3
2.85 g   KNO3
45 g      sucrose

volume up to 1500 ml
split into 3 x 500 ml

1)      OMS
No addition

2)      Zeatin
250 µl (1 mg/ml) zeatin, (0.5 mg/l zeatin final)

3) PCPA (parachlorophenoxy acetic acid) 250 µl (1 mg/ml) (0.5 mg/l PCPA final)

Each adjust to pH 5.7

Add 1 g Gelrite

Autoclave, pour medium #1, #3 into tall plates

Explant materials include seeds immature flower and possible stem or leaf tissue. Seeds should germinate on OMS and form multiple shoots on zeatin-containing medium. Immature flowers should form fruits in vitro on the medium containing PCPA. Germinating seeds will be used for subsequent experiments on Agrobacterium-mediated transformation using GFP.

Results 


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